A Guide to Commercial Multiplexing Platforms

Soluble analytes are among the most informative biomarkers in both research and diagnostics. Complex contexts, such as cancer, inflammation, or neurodegeneration, demand the simultaneous analysis of many biomarkers to reflect diverse and dynamic disease biology. This need is especially pronounced in clinical applications, where limited sample volumes make efficiency and cost-effectiveness essential. Multiplex immunoassays rise to these challenges by delivering higher throughput, richer data, and more comprehensive profiling from a single sample. Among their most exciting applications is biomarker discovery in liquid biopsies, such as in exploring blood plasma and serum proteomes. Here, we spotlight leading commercial platforms and offer guidance for researchers considering various avenues in multiplexing.

Bead-based multiplex immunoassays

High-plex bead-based multiplex immunoassays were pioneered by Luminex’s xMAP^® technology. In this platform, microscopic beads (microspheres) are coated with capture antibodies specific to target analytes, along with unique ratios of fluorescent dyes. This generates spectrally distinct signatures for distinguishing individual bead types. The beads are microscopic and low-density, and assay reactions proceed with fast binding kinetics inherent in solution-phase interactions. However, the solid-phase nature of the beads allows discrete quantification of each immunocomplex. This enables dedicated analyzers, equipped with multiple laser sources, to spectrally identify bead types as well as reporter fluorophores bound during analyte detection. The precise measurement of each bead–analyte complex enables true multiplexing, with the capacity to detect tens to hundreds of distinct analytes in a single sample.

"xMAP Technology really shines in scalability, sample volume, and speed. It can multiplex up to 500 analytes in a reaction, giving researchers a wealth of data from a single low-volume sample—and making it a great fit for analyzing precious samples," says Sherry Dunbar, Ph.D., Senior Director of Scientific Affairs Programs at Luminex. 

While Luminex offers several xMAP instrumentation options, it relies on a network of partners to develop and commercialize immunoassays. Several partners offer a variety of specialized kits, reagents, and complementary equipment. Notable examples include Bio-Plex^® (Bio-Rad), MILLIPLEX^® (MilliporeSigma), ProcartaPlex™ (Thermo Fisher Scientific), and Luminex^® Assays (R&D Systems).

Beyond xMAP, several other bead-based platforms provide alternative approaches to multiplexing, including EYRAplex (Mabtech AB), iQue Qbeads^® PlexScreen (Sartorius), LEGENDplex™ (BioLegend), EcoPlex™ (Elabscience), and Simoa^® bead-based assays (Quanterix). These systems share common design features—beads coated with analyte-specific antibodies and internally dyed for spectral distinction—while retaining the advantages of solution-phase reaction kinetics. Many can be analyzed on standard flow cytometers, offering flexible and accessible readout options, although Mabtech’s EYRA instrument instead employs confocal imaging and avoids fluidics entirely. Each platform also brings distinct strengths, ranging from high-throughput capacity and application-specific optimization to reduced cross-reactivity, customizable assay panels, and robust sensitivity.

Array-based multiplexing

High-plex immunoassays of multiple analytes are available beyond bead-based methods. Among these is the microarray format, in which distinct antibodies are immobilized on a solid support, typically a glass slide or nitrocellulose membrane, creating a two-dimensional array for parallel analyte detection. Although antibody microarrays have been in use for many years, they remain a robust option for large-scale proteomic screening. Antibody arrays from Creative Biolabs and RayBiotech feature profiling capabilities of over a thousand analytes, while Arrayjet’s Miniaturised Immunoassay platform extends coverage to over ten thousand targets. 

“If large panels of analytes need screening (100s-10,000s), microarray is the solution with the highest speed and throughput,” says Adam Buckle, Ph.D., Chief Scientific Officer at Arrayjet. “The miniaturised nature of the screen also drastically reduces materials usage and time. For example, a screen of 25,000 analytes could require ~130 microplates, but can be condensed onto just one microarray chip.”

Another approach to multiplexing is offered by planar array platforms, which print multiple discrete spots of capture antibodies onto the surface of wells in a microtiter plate. Each spot corresponds to a specific analyte, thereby expanding the multiplexing capacity of a conventional plate-based assay. Examples include Q-Plex™ Multiplex Assays (Quansys Biosciences and BosterBio), MULTI-SPOT^® and MULTI-ARRAY^® technologies (Meso Scale Discovery), and Simoa Planar Array Assays (Quanterix). These systems build on the familiarity and robustness of ELISA-style formats while conserving sample volume, increasing assay efficiency, and enabling the simultaneous quantification of multiple biomarkers within a single well.

Additional platforms for multiplexing

Beyond bead- and array-based systems, additional platforms for multiplexing leverage DNA barcoding to achieve high plex and sensitivity. In Olink’s Proximity Extension Assay (PEA), matched antibody pairs are conjugated to DNA oligonucleotides that, upon binding the same target analyte, come into proximity and are extended to form amplifiable DNA barcodes. The NULISA™ platform uses a similar principle, in which antibody pairs linked to unique DNA oligos generate analyte-specific barcodes when bound to their targets. In both systems, quantification is achieved through either real-time PCR or next-generation sequencing. These approaches are highly sensitive and scalable, enabling the simultaneous measurement of thousands of biomarkers with strong specificity and reproducibility.

Microfluidic technologies provide additional approaches for efficient multi-analyte analysis. These systems retain the core principles of ELISA-based immunoassays but process samples in parallel within automated microfluidic formats. The Ella™ and Simple Plex™ platform (ProteinSimple) uses preloaded microfluidic cartridges capable of quantifying up to eight analytes per run. In contrast, the Gyrolab^® system employs nanoliter-scale microfluidic CDs to miniaturize singleplex ELISAs and execute them in parallel, offering an alternative to conventional multiplexing. By integrating miniaturization with automation, both platforms generate high-quality data from minimal sample volumes while reducing hands-on time, pipetting variability, and plate-based incubation steps.

Considerations in choosing a platform

The commercial multiplexing landscape has grown increasingly diverse, with a wide range of specialized platforms available to researchers seeking higher throughput or broader analyte coverage. To help navigate these options, we spoke with experts from leading multiplex technology providers, who shared their insights on key considerations when selecting a platform.

“Start with the biology and sample—targets, expected concentrations, matrix, and available volume,” advises Vanitha Margan, Global Product Manager of Immunoassays at Bio-Rad. “Then compare platforms on sensitivity, dynamic range, multiplex capacity, time-to-result, throughput/automation, sample volume, reproducibility, and ease of use—along with total cost per usable data point. Also review cross-reactivity controls and standardization across lots/runs. Finally, match analysis software to your needs (quantitation, QC, automation). Keep it simple and deliberate—define your application and “must-have” criteria first, then shortlist platforms that meet them.”

Researchers should prioritize platforms that offer fit-for-purpose versatility, including configurable kits, premixed panels, or bulk formats tailored to different experimental requirements, advises Brooke Gilliam, Global Senior Product Manager of Immunoassay Platform Solutions at MilliporeSigma. “MILLIPLEX multiplex assays support relevant target models or disease-specific panels, making them ideal for translational and discovery research. High throughput and ease of use help streamline lab operations, and strong data quality and analysis tools, like Belysa^® Immunoassay Curve Fitting Software, ensure reliable, actionable results,” Gilliam adds.  

Experts from Mabtech* remind users to consider not only assay performance but also how an instrument will fit into daily lab routines. “Instruments that minimize downtime, maintenance, and training can have as much impact on productivity as sensitivity or panel size. With Mabtech EYRA, the absence of fluidics and the short read times mean the instrument is always ready when you are. This makes EYRA especially valuable in large studies or shared facilities where ease-of-use, minimal onboarding, and reproducibility across users are critical,” they add.

Long-term goals should be carefully considered, as multiplexing offers the greatest value when projects are expected to scale. “If researchers are starting with a few markers and might want to add more in the future—this is something we see very often in assay development—then incorporating multiplexing from the beginning gets them off to a great start and eliminates the headache of having to port a standard assay over to a multiplex assay later,” shares Dr. Dunbar from Luminex.

Finally, establishing a strong relationship with a supplier can be highly beneficial, providing both technical support and opportunities for collaboration. “From our perspective as a printing company, early-stage collaboration is essential to understand customers’ current and future needs. Providing guidance on panel design, assay optimisation, and integration into existing systems is crucial for long-term success and is a core differentiator of Arrayjet's end-to-end services,” adds Dr. Buckle from Arrayjet.

Table of multiplex platforms

Below are commercial multiplex platforms and assays from leading suppliers, organized by platform type. The table also indicates the maximum analyte capacity for each platform. For reference, links to relevant products are provided as available in our product directory. 

^*Tyler Sandberg, Product Manager of EYRAplex, ELISpot/FluoroSpot at Mabtech, and Berend Bassleer, Product Manager of Instruments at Mabtech.